Growth of recombinant equine herpesvirus 1 (EHV-1) replaced with passage-induced mutant gene 1 and gene 71 derived from an attenuated EHV-1 in cell cultures and in the lungs of mice

The relationship of passage-induced mutant genes 1 and 71 of an attenuated equine herpesvirus 1 (EHV-1) with virulence was analysed by constructing nine recombinant EHV-1 viruses by homologous recombination. Gene 1 or/and gene 71 of a virulent EHV-1 strain, HH1, was replaced by a mutant gene 1 or/and 71 of an attenuated HH1 strain, BK343, respectively. The beta-galactosidase gene of Escherichia coli was inserted within the gene 1 or 71 coding sequence of HH1 to inactivate the genes. Virus replications of these recombinant viruses in cell cultures were similar, but release of the gene 71-inactivated virus from infected cells was delayed compared to that of the other viruses. Plaque sizes of the recombinant viruses were similar to those of HH1, but those of BK343 were significantly smaller, indicating an effect of some unknown factor(s) on viral cell-to-cell spread. The growth abilities of the recombinant viruses with a mutant gene 1 or/and 71 in lungs of mice were similar to those of HH1, but those of gene 71-inactivated viruses were reduced to the level of BK343 and the titers were about 100-times lower than those of the other recombinant viruses. These results indicate that the mutant genes 1 and 71 of BK343 might not confer an attenuated nature to EHV-1.     

Mitochondrial DNA analysis of Nepalese domestic dwarf cattle Lulu*

Dwarf Lulu cattle, the only Bos Taurus type of cattle in Nepal, are raised under severe environments in the mountainous zone of that country. In the present study, the body measurement traits, cytogenetic and molecular genetic characteristics of the Lulu cattle are investigated. Blood samples were collected from 31 animals in four villages (altitudes 2590–3550 m) in the southern part of Mustang. The Lulu cattle had a normal karyotype with 2n = 60, XY or XX. Only one male examined had a large submetacentric X‐chromosome and a small submetacentric taurine type Y‐chromosome. The mitochodrial DNA (mtDNA) genotypes were analyzed by PCR mediated restriction fragment length polymorphisms, displacement (D)‐loop region PCR mediated single strand conformation polymorphisms, and D‐loop region sequences. Many base substitutions were found in the D‐loop region, suggesting that the Lulu cattle originated from at least 10 maternal lines. Three types of mtDNA from these cattle were found, the Bos taurus type (n = 23), the Bos indicus type (n = 6), and the Bos grunniens type (n = 2). In the village at the lowest altitude, four of the five cows were of the Bos indicus type. These results indicated that mtDNA types of the Lulu cattle mostly belong to Bos taurus, but have been hybridized with Bos indicus cattle in lower‐elevation regions in their maternal lineage.     

戊型肝炎病毒(HEV)猪间传播的动力学分析——对人类感染的潜在影响

【目的】戊型肝炎病毒(Hepatitis Evirus,HEV)感染是一种人畜共患病,而猪是HEV的病毒储库。在日本,HEV感染已成为猪的地方性猪病。阐明HEV在猪场中的详细传播机制,将有利于人们掌握特定年龄段猪群的感染状况,尤其是肉猪在屠宰前的感染状况。【结果】本文,作者重新分析了源于日本猪场的一个大型猪HEV血清阳性率数据集,以此评估HEV的感染力。结果表明,北海道、本州和九州三个地区猪场中的猪HEV感染力分别为3.45×10-2/d(95%的置信区间(下同):3.17～3.75×10-2/d)、2.68×10-2/d(2.28～3.14×10-2/d)和3.11×10-2/d(2.76～3.50×10-2/d)。利用作者建立的模型得到的估测结果表明,猪感染HEV的平均日龄在59.0～67.3d之间,其基本传染数R0按4.02～5.17的数量级递增。对特定年龄猪群在不同感染力下的敏感性分析显示,HEV感染力下降将提高猪感染HEV的年龄,而且会增加180日龄时散毒猪的数量。【结论】尽管作者的研究表明95%以上的猪是在150日龄前被感染的,但该模型显示感染力下降可以增加感染猪将HEV传播给人的风险。如果感染力开始下降,就有必要采取基本的应对策略(如在哺乳期末将未感染猪与感染猪分开)以最大限度地减少肥育阶段的HEV阳性猪数量。     

Proteome analysis of cerebrospinal fluid in healthy beagles and canine encephalitis

We performed proteomics analysis of the cerebrospinal fluid (CSF) of healthy dogs and dogs with meningoencephalitis of unknown etiology (MUE). By comparing two-dimensional electrophoreses (2DE), an upregulated spot was found in MUE dogs. This protein was identified as a neuron-specific enolase (NSE) by analysis with MALDI-TOF mass spectrometry. In comparing dot blots using an antibody against NSE, the NSE levels in the CSF of MUE dogs was significantly higher than that of the controls. NSE is a diagnostic marker of neuroendocrine tumors, brain injury and spinal cord trauma in humans. It seems that the NSE concentration in the CSF is increased by cellular destruction in canine encephalitis. Though elevation of NSE may not be specific in canine encephalitis because the NSE level was increased in other CNS diseases, further study including measurement with serum is necessary.     

Identification of weakly beta-hemolytic porcine spirochetes by biochemical reactions, PCR-based restriction fragment length polymorphism analysis and species-specific PCR

We examined the usefulness of PCR-based restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR combined with a newly devised rapid biochemical test using microplates for identifying weakly beta-hemolytic intestinal spirochetes (WBHIS) isolated from pigs. WBHIS strains showing atypical biochemical characteristics were decisively identified at the species level by PCR-RFLP and species-specific PCR. Identification of WBHIS at the species level in routine diagnostic work will certainly contribute to clarifying the pathogenicity of WBHIS.     

Bow variation in kiln-dried boxed-heart square timber of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) cultivars

To evaluate the bow variation in boxed-heart square timber of sugi (Cryptomeria japonica), bows from kiln-dried timber for five sugi cultivars with different longitudinal shrinkage trends were compared for two stem heights. Two general trends were observed, depending on the cultivar: (1) either the bow was larger at the lower than at the upper part of the stem, or (2) the bows at the lower and the upper parts of the stem were similar. In timber that had larger bow values, the gradients of longitudinal shrinkage were large across the radius and along the length of the timber. There was a positive relationship between the bow and longitudinal shrinkage. These results suggest that the bow variation between the timbers was caused by a variation in longitudinal shrinkage, which was affected by the microfibril angle. Furthermore, the bow was inversely proportional to the modulus of elasticity, which suggests that timber with a low modulus of elasticity is susceptible to a large bow due to large longitudinal shrinkage.     

Shear strengths of wood measured by various short beam shear test methods

We conducted three types of short beam shear tests of western hemlock (Tsuga heterophylla Sarg.) under various span/depth ratios, and examined whether the maximum shear stress was used as the shear strength. The following results were obtained. (1) In the short beam shear tests under the three-point loading method, it was difficult to have the specimen failing by horizontal shear. We thought that this method should not be recommended for determining the shear strength of wood. (2) In the short beam shear tests under the asymmetric four-point loading of the specimen with a rectangular cross-section, the failure caused by horizontal shear occurred under some span/depth ratio range. Nevertheless, this range was dependent on the specimen geometry and was quite restricted. We therefore think that this method should not be recommended for determining the shear strength of wood. (3) In the short beam shear tests under the asymmetric four-point loading of the I-shaped specimen, failure caused by horizontal shear occurred under the span/depth ratio range wider than that applicable for the asymmetric four-point loading of the specimen with a rectangular cross-section. The maximum shear stress was stable in a certain span/depth ratio range and the value of the maximum shear stress is effective as a parameter for comparing the shearing strength of materials with each other.